Methods, compositions, and kits for rare allele detection

摘要

Methods are provided for nucleic acid analysis. In an illustrative method, Snapback-ARMS primers are used to amplify preferentially a target nucleic acid that is present in a low allele fraction. In another embodiment, tailed primers are used to identify the preferentially amplified allele.

主权项

1. A method for detecting a minor allele in a sample comprising,
amplifying DNA comprising the minor allele and a major allele, wherein the minor allele is present in a lower concentration than the major allele, and the minor allele differs from the major allele by a nucleotide, wherein the amplifying step is performed using
a pair of primers comprising an ARMS primer, wherein the ARMS primer is a Snapback ARMS primer, and a second primer, wherein a 3′ terminal nucleotide of the ARMS primer is complementary to the nucleotide of the minor allele and is not complementary to the nucleotide of the major allele, anda probe, wherein the probe is a probe element located as a 5′ end of the ARMS primer, wherein the probe has a nucleotide that is complementary to the nucleotide of the major allele and is not complementary to the minor allele, and detecting the minor allele using the probe under annealing or melting conditions.